Adipocytes from New Zealand Obese Mice Exhibit Aberrant Proinflammatory Reactivity to the Stress Signal Heat Shock Protein 60

Figure 3

Identification of Hsp60 binding regions by Hsp60 epitope-specific antibodies. Increasing concentrations (0–25 μg/mL) of antibodies directed against distinct aminoacid (aa) regions of the Hsp60 molecule were incubated with 100 nM Dylight649-labeled Hsp60 (Hsp60*) prior addition to C57BL/6J, NOD, or NZO mouse-derived preadipocytes. The resulting fluorescence signals of the cells were determined by FACS analysis. The data show residual Hsp60* binding to C57BL/6J (light grey bars), NOD (dark grey bars), and NZO mouse-derived cells (black bars) as percent of the binding of Hsp60* to cells cultivated in medium (Med.) in the absence of antibodies (set as 100%, white bar). The data represent means + SEM of three independent experiments. *𝑃<0.05; **𝑃<0.01; ***𝑃<0.001 compared to the binding of Hsp60* alone.